Hitoshi Nochi

Modulation of Hepatic Granulomatous Responses by Transgene Expression of DAP12 or TREM-1-Ig Molecules

DAP12 (also known as KARAP) is a novel ITAM-bearing transmembrane adapter molecule that is expressed on the cell surface of natural killer cells, monocytes, dendritic cells, and macrophages. Several myeloid cell-specific DAP12-associating receptors, such as TREM receptor family, SIRP-beta1, and MDL-1 have been identified. The in vivo function of DAP12 and its associating molecules in inflammation has remained primarily unknown. To investigate DAP12 signaling during chronic inflammation, we constructed two adenoviral gene transfer vectors to express FLAG/DAP12 (Ad-FDAP12) and the extracellular domain of mouse TREM-1 and the Fc portion of human IgG1 (Ad-TREM-1 Ig), respectively, and observed their modulatory activities in a mouse model of hepatic granulomatous inflammation elicited by zymosan A. Mice were injected with zymosan A intravenously and 24 hours after zymosan A, they were injected with Ad-FDAP12 or Ad-TREM-1 Ig. Zymosan A-induced hepatic granuloma formation peaked at day 7 and markedly declined by day 10. Although adenoviral-mediated DAP12 gene transfer did not enhance granuloma formation by day 7, it sustained and enhanced granuloma formation beyond day 7. However, an anti-FLAG monoclonal antibody used to potentiate the signaling of adenoviral-derived DAP12, enhanced granuloma formation at day 7. In sharp contrast to the effect by Ad-FDAP12, transgene expression in the liver of soluble form of extracellular domain of TREM-1 as an antagonist of DAP12 signaling, remarkably inhibited zymosan A-induced granuloma formation at all time points examined. Our findings thus suggest that both DAP12 and TREM-1 are involved in the development of granulomatous responses in the liver.

Identification of metallopanstimulin-1 as a member of a tumor associated antigen in patients with breast cancer

The immunological screening of breast cancer was performed with IgG autoantibodies by the serological analysis of recombinant cDNA expression library methods to explore novel tumor associated antigens. We have focused on a small zinc finger protein metallopanstimulin-1 (MPS-1). MPS-1 mRNA was ubiquitously expressed in normal human tissues except the brain and the placenta. In Western blot analysis, MPS-1 was easily detected strongly in actively proliferating cells and three breast cancer cell lines. In the tissue the protein of MPS-1 in cancer cells was more abundant than that of surrounding normal cells. Screening of tissue specimens by immunohistochemistry revealed 50.4% positive for MPS-1 in 125 cancer patients. These data suggest that MPS-1 could be applicable to the immunotherapy of breast cancer.

Acute rhabdomyolysis of the soleus muscle induced by a lightning strike: magnetic resonance and scintigraphic findings.

Among natural disasters, a lightning strike is a rare but potentially life-threatening phenomenon. If victims survive a cardiac arrest due to instantaneous passage of an exceptionally high voltage electric charge through the whole body, they may be afflicted with various complications such as muscle necrosis resulting in acute renal failure. In this article, we report a case of a 54-year-old man with acute rhabdomyolysis of the left soleus muscle associated with a lightning strike. T2-weighted and short-tau inversion recovery MR images showed a high signal intensity in the left soleus muscle. A whole-body bone scintigram showed abnormal uptakes in the left soleus muscle and the dorsal aspect of the left foot. MR and scintigraphic evaluations were very useful in depicting the site and extent of muscle damage. Since the patient showed a surprisingly high level of serum creatine kinase, the added information was very valuable for determining the patient’s management.

Alloreactivity and immunosuppressive properties of articular chondrocytes from osteoarthritic cartilage.

Purpose To determine whether articular chondrocytes derived from osteoarthritic knee joints could evoke alloreactive proliferation of peripheral blood mononuclear cells (PBMC) and inhibit mitogenic activity of polyclonally activated CD4+ major histocompatibility complex (MHC) class II–restricted T cells in vitro. Methods Osteoarthritic cartilages of 17 patients aged 61 to 85 years were harvested during total knee arthroplasty. Chondrocytes were cultured for experiments. PBMCs, CD4+ T cells, CD8+ T cells, and CD14+ monocytes from healthy subjects were also used. To investigate the allogeneic response and immunosuppressive properties of chondrocytes, assays for one-way mixed lymphocyte reaction (MLR), apoptosis, activated CD4+ T-cell proliferation, and cytotoxic CD8+ T-cells were performed. Chondrocyte cell-surface antigens were examined using flow cytometry. Results Chondrocytes failed to trigger an allogeneic PBMC reaction and did not induce apoptosis of allogeneic PBMCs in the MLR assay. Chondrocytes inhibited the proliferation of polyclonally activated CD4+ T cells via cell-cell contact and escaped the allogeneic cytotoxic reactivity of CD8+ T cells. Chondrocytes expressed MHC class I but not MHC class II molecules or B7-1/-2-positive co-stimulatory molecules. Conclusion Chondrocytes from osteoarthritic knees in older patients exhibited similar immunomodulatory properties in vitro to those in juveniles or adults.